{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T14:58:51Z","timestamp":1790002731212,"version":"4.0.1"},"reference-count":41,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2019,3,20]],"date-time":"2019-03-20T00:00:00Z","timestamp":1553040000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"publisher","award":["BB\/K002465\/1"],"award-info":[{"award-number":["BB\/K002465\/1"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012065","name":"Houghton Trust","doi-asserted-by":"publisher","award":["Small project grant to ES Giotis"],"award-info":[{"award-number":["Small project grant to ES Giotis"]}],"id":[{"id":"10.13039\/100012065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>The discovery of mammalian pluripotent embryonic stem cells (ESC) has revolutionised cell research and regenerative medicine. More recently discovered chicken ESC (cESC), though less intensively studied, are increasingly popular as vaccine substrates due to a dearth of avian cell lines. Information on the comparative performance of cESC with common vaccine viruses is limited. Using RNA-sequencing, we compared cESC transcriptional programmes elicited by stimulation with chicken type I interferon or infection with vaccine viruses routinely propagated in primary chicken embryo fibroblasts (CEF). We used poxviruses (fowlpox virus (FWPV) FP9, canarypox virus (CNPV), and modified vaccinia virus Ankara (MVA)) and a birnavirus (infectious bursal disease virus (IBDV) PBG98). Interferon-stimulated genes (ISGs) were induced in cESC to levels comparable to those in CEF and immortalised chicken fibroblast DF-1 cells. cESC are permissive (with distinct host transcriptional responses) to MVA, FP9, and CNPV but, surprisingly, not to PBG98. MVA, CNPV, and FP9 suppressed innate immune responses, while PBG98 induced a subset of ISGs. Dysregulation of signalling pathways (i.e., NF\u03baB, TRAF) was observed, which might affect immune responses and viral replication. In conclusion, we show that cESC are an attractive alternative substrate to study and propagate poxvirus recombinant vaccine vectors.<\/jats:p>","DOI":"10.3390\/genes10030237","type":"journal-article","created":{"date-parts":[[2019,3,21]],"date-time":"2019-03-21T04:11:56Z","timestamp":1553141516000},"page":"237","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Chicken Embryonic-Stem Cells Are Permissive to Poxvirus Recombinant Vaccine Vectors"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8908-7824","authenticated-orcid":false,"given":"Efstathios S.","family":"Giotis","sequence":"first","affiliation":[{"name":"Section of Virology, Department of Medicine, St Mary\u2019s Campus, Imperial College London, Norfolk Place, London W2 1PG, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillaume","family":"Montillet","sequence":"additional","affiliation":[{"name":"Univ Lyon, Universit\u00e9 Lyon 1, INSERM, INRA, Stem Cell and Brain Research Institute, U1208, USC1361 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bertrand","family":"Pain","sequence":"additional","affiliation":[{"name":"Univ Lyon, Universit\u00e9 Lyon 1, INSERM, INRA, Stem Cell and Brain Research Institute, U1208, USC1361 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0050-4167","authenticated-orcid":false,"given":"Michael A.","family":"Skinner","sequence":"additional","affiliation":[{"name":"Section of Virology, Department of Medicine, St Mary\u2019s Campus, Imperial College London, Norfolk Place, London W2 1PG, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,3,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1002\/biot.201400388","article-title":"Designing cell lines for viral vaccine production: Where do we stand?","volume":"10","author":"Genzel","year":"2015","journal-title":"Biotechnol. J."},{"key":"ref_2","first-page":"419","article-title":"The avian EB66\u00ae cell line, application to vaccines, and therapeutic protein production","volume":"64","author":"Brown","year":"2010","journal-title":"PDA J. Pharm. Sci. Technol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"6078","DOI":"10.1016\/j.vaccine.2015.09.022","article-title":"A clinical phase I study of an EB66 cell-derived H5N1 pandemic vaccine adjuvanted with AS03","volume":"33","author":"Naruse","year":"2015","journal-title":"Vaccine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"405","DOI":"10.4161\/mabs.12350","article-title":"EB66 cell line, a duck embryonic stem cell-derived substrate for the industrial production of therapeutic monoclonal antibodies with enhanced ADCC activity","volume":"2","author":"Olivier","year":"2010","journal-title":"MAbs"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/j.vaccine.2008.11.066","article-title":"An avian cell line designed for production of highly attenuated viruses","volume":"27","author":"Jordan","year":"2009","journal-title":"Vaccine"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2339","DOI":"10.1242\/dev.122.8.2339","article-title":"Long-term in vitro culture and characterisation of avian embryonic stem cells with multiple morphogenetic potentialities","volume":"122","author":"Pain","year":"1996","journal-title":"Development"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1111\/j.1440-169X.2009.01152.x","article-title":"Chicken embryonic stem cells as a non-mammalian embryonic stem cell model","volume":"52","author":"Lavial","year":"2010","journal-title":"Dev. Growth Differ."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1007\/978-1-62703-628-3_11","article-title":"Chicken embryonic stem cells: Establishment and characterization","volume":"1074","author":"Aubel","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1387\/ijdb.170322bp","article-title":"Pluripotency in avian species","volume":"62","author":"Pain","year":"2018","journal-title":"Int. J. Dev. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1093\/infdis\/jiv091","article-title":"Immunogenicity and safety of an EB66 cell-culture-derived Influenza A\/Indonesia\/5\/2005(H5N1) AS03-adjuvanted vaccine: A phase 1 randomized trial","volume":"212","author":"Schuind","year":"2015","journal-title":"J. Infect. Dis."},{"key":"ref_11","unstructured":"Mehtali, M., Champion-Arnaud, P., and Arnaud, L. (2015). Production of viral vaccines in suspension on avian embryonic derived stem cell lines. (No. 9040296), U.S. Patent."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.scr.2014.11.005","article-title":"Transcriptome analysis of chicken ES, blastodermal and germ cells reveals that chick ES cells are equivalent to mouse ES cells rather than EpiSC","volume":"14","author":"Jean","year":"2015","journal-title":"Stem Cell Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1272","DOI":"10.1016\/j.stemcr.2018.09.005","article-title":"NANOG is required for the long-term establishment of avian somatic reprogrammed cells","volume":"11","author":"Fuet","year":"2018","journal-title":"Stem Cell Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1159\/000016701","article-title":"Chicken embryonic stem cells and transgenic strategies","volume":"165","author":"Pain","year":"1999","journal-title":"Cells Tissues Org."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Giotis, E.S., and Skinner, M.A. (2018). Spotlight on avian pathology: Fowlpox virus. Avian Pathol., 87\u201390.","DOI":"10.1080\/03079457.2018.1554893"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1099\/vir.0.19568-0","article-title":"Comparison of the genome sequence of FP9, an attenuated, tissue culture-adapted European strain of Fowlpox virus, with those of virulent American and European viruses","volume":"85","author":"Laidlaw","year":"2004","journal-title":"J. Gen. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2191","DOI":"10.1099\/vir.0.81738-0","article-title":"Avipoxvirus phylogenetics: Identification of a PCR length polymorphism that discriminates between the two major clades","volume":"87","author":"Jarmin","year":"2006","journal-title":"J. Gen. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2386","DOI":"10.1055\/s-0028-1108143","article-title":"MVA vaccination against smallpox: Clinical tests with an attenuated live vaccinia virus strain (MVA) (author\u2019s transl)","volume":"99","author":"Stickl","year":"1974","journal-title":"Dtsch. Med. Wochenschr."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0168-1702(95)01253-2","article-title":"Coding sequences of both genome segments of a European \u2018very virulent\u2019 infectious bursal disease virus","volume":"40","author":"Brown","year":"1996","journal-title":"Virus Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1099\/0022-1317-79-5-1047","article-title":"Expression of ORF A1 of infectious bursal disease virus results in the formation of virus-like particles","volume":"79","author":"Risco","year":"1998","journal-title":"J. Gen. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5028","DOI":"10.1128\/JVI.02738-12","article-title":"Genetic screen of a library of chimeric poxviruses identifies an ankyrin repeat protein involved in resistance to the avian type I interferon response","volume":"87","author":"Buttigieg","year":"2013","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5041","DOI":"10.1128\/JVI.02736-12","article-title":"Genetic screen of a mutant poxvirus library identifies an ankyrin repeat protein involved in blocking induction of avian type I interferon","volume":"87","author":"Laidlaw","year":"2013","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Giotis, E.S., Scott, A., Rothwell, L., Hu, T., Talbot, R., Todd, D., Burt, D.W., Glass, E.J., and Kaiser, P. (2018). Chicken anaemia virus evades host immune responses in transformed lymphocytes. J. Gen. Virol.","DOI":"10.1099\/jgv.0.001011"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1859","DOI":"10.1091\/mbc.10.6.1859","article-title":"Dynamics of gene expression revealed by comparison of serial analysis of gene expression transcript profiles from yeast grown on two different carbon sources","volume":"10","author":"Kal","year":"1999","journal-title":"Mol. Biol. Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1186\/s13567-016-0363-8","article-title":"Chicken interferome: Avian interferon-stimulated genes identified by microarray and RNA-seq of primary chick embryo fibroblasts treated with a chicken type I interferon (IFN-\u03b1)","volume":"47","author":"Giotis","year":"2016","journal-title":"Vet. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.1101\/gr.400202","article-title":"The automatic detection of homologous regions (ADHoRe) and its application to microcolinearity between Arabidopsis and rice","volume":"12","author":"Vandepoele","year":"2002","journal-title":"Genome Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/978-1-4939-7216-6_14","article-title":"Chicken induced pluripotent stem cells: Establishment and characterization","volume":"1650","author":"Fuet","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1093\/humupd\/dml041","article-title":"Differences between human embryonic stem cell lines","volume":"13","author":"Allegrucci","year":"2007","journal-title":"Hum. Reprod. Update"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"17485","DOI":"10.1038\/s41598-017-17730-2","article-title":"Constitutively elevated levels of SOCS1 suppress innate responses in DF-1 immortalised chicken fibroblast cells","volume":"7","author":"Giotis","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1038\/nri3581","article-title":"Regulation of type i interferon responses","volume":"14","author":"Ivashkiv","year":"2014","journal-title":"Nat. Rev. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1230","DOI":"10.1038\/nature06403","article-title":"Nanog safeguards pluripotency and mediates germline development","volume":"450","author":"Chambers","year":"2007","journal-title":"Nature"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1381","DOI":"10.1292\/jvms.11-0517","article-title":"Biological effects of chicken type III interferon on expression of interferon-stimulated genes in chickens: Comparison with type I and type II interferons","volume":"74","author":"Masuda","year":"2012","journal-title":"J. Vet. Med. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Qu, H., Yang, L., Meng, S., Xu, L., Bi, Y., Jia, X., Li, J., Sun, L., and Liu, W. (2013). The differential antiviral activities of chicken interferon \u03b1 (ChIFN-\u03b1) and ChIFN-\u03b2 are related to distinct interferon-stimulated gene expression. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0059307"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.vetimm.2011.05.031","article-title":"Identifying innate immune pathways of the chicken may lead to new antiviral therapies","volume":"148","author":"Karpala","year":"2012","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Liem, J., and Liu, J. (2016). Stress beyond translation: Poxviruses and more. Viruses, 8.","DOI":"10.3390\/v8060169"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3514","DOI":"10.1128\/MCB.26.9.3514-3526.2006","article-title":"Differential regulation of the transcriptional activities of hypoxia-inducible factor 1 \u03b1 (HIF-1\u03b1) and HIF-2\u03b1 in stem cells","volume":"26","author":"Hu","year":"2006","journal-title":"Mol. Cell. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1101\/gad.1399906","article-title":"Hif-2\u03b1 regulates Oct-4: Effects of hypoxia on stem cell function, embryonic development, and tumor growth","volume":"20","author":"Covello","year":"2006","journal-title":"Genes Dev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.3181\/0709-RM-263","article-title":"Activation of hypoxic response in human embryonic stem cell-derived embryoid bodies","volume":"233","author":"Cameron","year":"2008","journal-title":"Exp. Biol. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/1750-2187-8-7","article-title":"TRAF molecules in cell signaling and in human diseases","volume":"8","author":"Xie","year":"2013","journal-title":"J. Mol. Signal"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2918","DOI":"10.1099\/jgv.0.000979","article-title":"Differential gene expression in chicken primary B cells infected ex vivo with attenuated and very virulent strains of infectious bursal disease virus (IBDV)","volume":"98","author":"Dulwich","year":"2017","journal-title":"J. Gen. Virol."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/10\/3\/237\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:39:24Z","timestamp":1760186364000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/10\/3\/237"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,3,20]]},"references-count":41,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2019,3]]}},"alternative-id":["genes10030237"],"URL":"https:\/\/doi.org\/10.3390\/genes10030237","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,3,20]]}}}